Double staining of proteoglycans and the HNK-1 carbohydrate epitope in pseudoexfoliation material.
OBJECTIVE
We combined the techniques of both immunogold and cupromeronic-blue staining with electron microscopy to investigate the possible relationship between proteoglycans and the HNK-1 carbohydrate epitope in pseudoexfoliation material.


METHODS
The anterior segment was obtained from an enucleated eye with pseudoexfoliation syndrome. After cupromeronic-blue staining, following the method of Scott, specimens were embedded in L. R. White resin. Ultrathin sections were incubated successively in primary monoclonal antibodies, and 10 nm gold-conjugated secondary antibody.


RESULTS
The present study showed simultaneous presence of the HNK-1 epitope and proteoglycans associated with zonular fibrils and pseudoexfoliation material on the posterior surface of the iris. The HNK-1 epitope was immunolocalized to the periphery of the pseudoexfoliation fibers, while cupromeronic blue-positive filaments bridged adjacent pseudoexfoliation fibers.


CONCLUSIONS
Proteoglycans and the HNK-1 epitope might play a substantial role in the formation of pseudoexfoliation material via their adhesive function. Proteoglycans probably do not directly bear the HNK-1 epitope in pseudoexfoliation material.
Kubota
T
T
Department of Ophthalmology, Faculty of Medicine, Kyushu University, Fukuoka, Japan. kubota@eye.med.kyushu-u.ac.jp
Khalil
A
A
Tawara
A
A
Zhang
X
X
Inomata
H
H
D000368:Aged; D000869:Anterior Eye Segment / Q000378:metabolism* / Q000648:ultrastructure; D018959:CD57 Antigens / Q000378:metabolism*; D004396:Coloring Agents; D000939:Epitopes / Q000378:metabolism*; D017889:Exfoliation Syndrome / Q000378:metabolism* / Q000473:pathology; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007211:Indoles; D016253:Microscopy, Immunoelectron; D009942:Organometallic Compounds; D011509:Proteoglycans / Q000378:metabolism*
D018959:CD57 Antigens; D004396:Coloring Agents; D000939:Epitopes; D007211:Indoles; D009942:Organometallic Compounds; D011509:Proteoglycans; C015445:copper phthalocyanine